ITS1 gene sequences from human gut
Sequencing was performed at the Centre for Proteomic and Genomic Research (CPGR) in Cape Town, on the Illumina MiSeq platform. Targeted sequencing was performed using the Illumina Fungal Metagenomic Demonstrated Sequencing Protocol and internal transcribed spacer 1 (ITS1) primer pool [https://emea.support.illumina.com/content/dam/illumina-support/documents/documentation/chemistry_documentation/metagenomic/fungal-metagenomic-demonstrated-protocol-1000000064940-01.pdf.]. The amplicon PCR was adapted by increasing the number of cycles to 35 and the forward and reverse primer pools were added at concentrations of 2 ?M and 1 ?M, respectively. The MiSeq Reagent v3 Kit (600 cycles) was used to generate sequencing libraries for 2??200 bp paired-end read sequencing. Sequencing libraries were spiked with 10% of a 5 pM PhiX sequencing control. Libraries were purified using AMPure XP beads (Beckman Coulter). All PCR reactions utilized KAPA HiFi HotStart ReadyMix (Roche). Prior to pooling, dsDNA was measured in each library using the QuantiFluor dsDNA System (Promega), and library sizes were verified using the D1000 (Agilent) the 4200 TapeStation System.